The objective of this four year renewal program is to complete the development of enhanced serological and virological test systems for B virus (Herpesvirus simiae) as well as other simian herpesviruses, to unambiguously identify B virus infection in humans, macaques, or other exposed nonhuman primates. We will exploit structural and functional properties of B virus, and closely related herpesviruses to complete development of a multifaceted diagnostic system. We propose development and trial of the following assay systems: (1) a rapid enzyme-immunoassay for serological identification of B virus specific antibodies using recombinant B virus components to replace currently required B virus propagation, (2) a rapid antigen detection system to identify the presence of replication competent B virus present in human and nonhuman primate tissues (lesional or nonlesional sites by polymerase chain reaction (PCR), and (3) molecular analysis using in situ hybridization (ISH) coupled with PCR techniques to identify primates with potential for reactivation of latent B virus. To accomplish the development of these systems we propose the following specific aims; (1) utilize the identified polypeptides/glycoproteins of B virus to which human (n=520) and macaque (n=greater than 1000) virus hosts have been shown to direct a specific humoral response (IgM and IgG), (2) identify B virus specific sequences, particularly those identified which code for unique, specific epitopes in these B virus polypeptide(s)/glycoprotein(s), (3) clone these sequences for diagnostic use in PCR and ISH, (4) express these sequences in a high efficiency Autographa californica vector system, (5) develop rapid assay(s) systems using these recombinant expression products, and (6) produce polyclonal monospecific antibodies and monoclonal antibody reagents to develop antigen capture assays for rapid identification of B virus in field use. The results of the research proposed in this renewal application will provide the basis for implementing the safest possible operating standards for NIH and other federally supported macaque resource colonies and their personnel. Adoption of these standards will enhance management, prevention and control of B virus infections in humans and other primate species. Development of these diagnostic systems is especially important in preventing further zoonotic disease in humans now that macaque usage is increasing in response to the applicability of this species in AIDS research.